In the last years, there has been a growing interest in the recovery of bioactive compounds from natural sources for the development of novel functional foods. In this study, natural extracts, previously obtained by fractioned high pressure extraction from "Saco" cherry culls, were characterized in terms of anticancer activity. The product derived from CO2:EtOH (90:10, v/v) extraction after a pre-treatment of raw material with supercritical CO2 during 1 h, exhibited the highest antiproliferative activity in human HT29 colon cancer cells. In addition, when compared with doxorubicin, cherry extracts induced cell cycle arrest in a different cell cycle checkpoint. In order to obtain extracts with enhanced antiproliferative activity, the extraction process was further explored. Using highly pure CO2 and EtOH 96% the inhibition of cancer cell growth was significantly enhanced by 16-fold. In addition, the incorporation of a conventional extraction step with MetOH or EtOH:H2O (50:50, v/v) prior to fractioned process allowed to obtain cherry extracts more concentrated in antiproliferative compounds. Perillyl alcohol present in cherry extracts was pointed to be one of the major responsible for anticancer properties of cherry extracts. (C) 2010 Elsevier B.V. All rights reserved.